Registration Dossier

Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
15 May 2017 - 19 May 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report Date:
2017

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Deviations:
no
Qualifier:
according to
Guideline:
ISO 7346-1 (Determination of the Acute Lethal Toxicity of Substances to a Freshwater Fish [Brachydanio rerio Hamilton-Buchanan (Teleostei, Cyprinidae)] - Part 1: Static Method)
Deviations:
no
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Perfluorpropylvinylether (PPVE), MTDID 16437
- Substance type: pure active substance
- Physical state: liquid
- Analytical purity: 98.5%
- Storage condition of test material: At room temperature in the dark under nitrogen
- Other: Flush container with nitrogen after handling
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 115B1065A
- Physical state: clear colorless liquid
- Analytical purity: 98.7%
- Expiration date of the lot/batch: 26 Nov 2018

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature in the dark under nitrogen
- Stability under test conditions: stable

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Concentrations: negative control and saturated solution (limit test)
- Volume: Control: 10 mL, Saturated solution: 0.30 mL
- Sampling method: Duplicate samples for analysis were taken from one of the three replicate bottles from both test groups at the start of the test and at 96 hours. In addition, a quality control sample of 0.15 mL was taken from the saturated solution at the start and the end of the test period.
- Sample storage conditions before analysis: Samples were analyzed on the day of sampling.

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The test substance is extremely volatile. Therefore, 3.3L screw-top glass flasks were completely filled with medium and closed with a septum-sealed screw cap and parafilm. PPVE (21.25 µL) was then injected through the septum with gas-tight syringe. Flasks were rotated slowly for five days, and then allowed to settle for two days.
- Controls: Blank only
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): Test solutions were clear and colorless, with a minimum of visible droplets of undissolved test substance.

Test organisms

Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: Zebra-fish
- Source: Zodiac, proeffac, De Haar Vissen, Wageningen University and Research Centre, The Netherlands
- Length at study initiation: 1.8 ± 0.2 cm
- Weight at study initiation: 0.10 ± 0.03 g
- Feeding during test: no

ACCLIMATION
- Acclimation period: at least 12 days after delivery
- Acclimation conditions (same as test or not): Adjusted ISO medium, formulated using RO-water (tapwater purified by reverse osmosis; GEON Waterbehandeling, Berkel-Enschot, The Netherlands) with the following composition:
CaCl2.2H2O 211.5 mg/L
MgSO4.7H2O 88.8 mg/L
NaHCO3 46.7 mg/L
KCl 4.2 mg/L
- Type and amount of food during acclimation: pelleted fish food (Essence (300-500 um), Coppens International bv, Helmond, The Netherlands)
- Feeding frequency during acclimation: daily until 24 hours prior to the start of the test
- Health during acclimation (any mortality observed): than 5% mortality in the seven days prior to the test

Study design

Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h

Test conditions

Hardness:
180 mg CaCO3/L
Test temperature:
21.8 - 22.0 °C
pH:
7.1 - 8.0
Dissolved oxygen:
4.9 - 9.2 mg/L
Nominal and measured concentrations:
Nominal: 10 mg/L
Measured: 0.52 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: 3.3 liter glass flasks
- Type: closed
- Fill volume: 3.3 liters
- Aeration: no
- No. of organisms per vessel: 3
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 3
- Biomass loading rate: 0.09 g fish/liter

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: ISO medium prepared using reverse-osmosis purified water (GEON Waterbehandeling, Berkel-Enschot, The Netherlands)
- Intervals of water quality measurement: Daily in all vessels with surviving fish, beginning at the start of the test (day 0)

OTHER TEST CONDITIONS
- Adjustment of pH: None
- Photoperiod:16 hours light; 8 hours dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Mortality and/or severely distressed fish at 3.25, 24, 48, 72, and 96 hours.

TEST CONCENTRATIONS
- Test concentrations: 10 mg/L
Reference substance (positive control):
yes
Remarks:
pentachlorophenol (PCP)

Results and discussion

Effect concentrations
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 0.52 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: 0.52 mg/L was considered the maximum soluble concentration in test medium. Test conducted at a loading rate of 10 mg/L.
Details on results:
- Behavioural abnormalities: Two out of the nine fish exposed to the test item treated solutions died during the 96-hour test period. All fish were observed to swim slowly from 72 hours onwards when compared to the fish in the control group.
- Mortality of control: None
- Other adverse effects control: None
- Effect concentrations exceeding solubility of substance in test medium: Yes

-Other: In a preliminary test, no mortality or clinic effects were observed until day 2, while all fish exposed to the saturated 10 mg/L loading were found dead on day 3, i.e. after 72 hours of exposure. The study was interrupted and the physical parameters oxygen, pH and temperature were measured. Results of pH and temperature showed values within acceptable normal criteria for these parameters, however oxygen concentration in the saturated solution had decreased from 8.5 mg/L at the start to 0.7 mg/L after 72 hours. The oxygen concentration in the control had also decreased but less dramatically from 8.6 mg/L at the start to 2.4 mg/L after 72 hours. It was therefore expected that fish in the saturated solution had died due to a lack of oxygen.
Results with reference substance (positive control):
- Results with reference substance valid? yes
- Mortality: yes
- LC50: 0.15 mg/L
- Other: The sensitivity of zebra-fish originating from the present batch for PCP falls within the expected range of sensitivities generally observed for a similar species (carp) during the past years. Reference test done 14 days prior to limit test.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Remarks:
No control mortality, DO > 60% of saturation, results are based on measured concentrations.
Conclusions:
The 96-hour LC50 of PPVE to Danio rerio was >0.52 mg/L which was considered the maximum soluble concentration in the test medium (OECD 203, EU Method C.1 and ISO 7346-1).
Executive summary:

The 96-hour LC50 of PPVE to Danio rerio was examined in a static test conducted according to OECD 203, EU Method C.1, and ISO 7346-1. Test solutions were prepared as the control (0 mg/L) and at 10 mg/L. Analytically determined concentrations were < LOQ, and 0.52 mg/L, which was considered the maximum soluble concentration in the test medium. At 0.52 mg/L, cumulatively two of nine fish died by 96 hours. The 96-hour LC50 is > 0.52 mg/L.

The test was conducted according to internationally accepted test guidelines and was GLP compliant. It is reliable without restriction and suitable for Risk Assessment, Classification and Labeling, and PBT Analysis.